Targeting cell death in vivo in experimental traumatic brain injury by a novel molecular probe
Targeting cell death in vivo in experimental traumatic brain injury by a novel molecular probe
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DOI：
10.1089/neu.2007.0341
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发表时间：
2008-06-01
影响因子：
4.2
通讯作者：
Ziv, Ilan
中科院分区：
文献类型：
--
作者：
Reshef, Ayelet;Shirvan, Anat;Ziv, Ilan
创伤性脑损伤（TBI）仍然是神经和神经外科实践中常见的主要挑战。细胞凋亡可能在创伤性损伤诱导的脑组织损伤中发挥作用，因此其检测和抑制可能会促进患者护理。DDC（N，N '-didansyl-L-cystine）是一种新型的凋亡荧光探针。我们现在报告DDC在实验性TBI中的性能。采用小鼠闭合性颅脑损伤模型。DDC在体内静脉内给药。两小时后，处死动物，并对脑组织进行荧光显微镜检查，以评估DDC摄取，与通过TUNEL和半胱天冬酶底物进行的细胞凋亡的组织病理学评估相关，并且还与通过神经严重程度评分（NSS）评估的神经功能缺损相关。DDC的选择性摄取观察到在原发部位的损伤，也在远程网站。摄取在细胞水平，DDC在细胞质中蓄积。通过检测特征性凋亡DNA片段化（阳性TUNEL染色）和检测活化的半胱天冬酶，证实显示DDC摄取的细胞为凋亡细胞。DDC荧光染色的受损区域与神经元缺损的严重程度相关。我们的研究证实了细胞凋亡在TBI中的作用，并提出DDC作为其选择性靶向和体内检测的有用工具。在DDC的未来放射性标记之后，这种细胞凋亡成像可以通过允许实时评估组织损伤的程度、评估新的治疗策略和优化个体患者的治疗来促进头部损伤患者的护理。
Traumatic brain injury (TBI) remains a frequent and major challenge in neurological and neurosurgical practice. Apoptosis may play a role in cerebral tissue damage induced by the traumatic insult, and thus its detection and inhibition may advance patient care. DDC (N, N'-didansyl-L-cystine) is a novel fluorescent probe for detection of apoptotic cells. We now report on the performance of DDC in experimental TBI. Closed head injury was induced in mice by weight-drop. DDC was administered intravenously in vivo. Two hours later, animals were sacrificed, and brain tissue was subjected to fluorescent microcopy, for assessment of DDC uptake, in correlation with histopathological assessment of apoptosis by TUNEL and caspase substrates, and also in correlation with the neurological deficits, as assessed by Neurological Severity Score (NSS). Selective uptake of DDC was observed at the primary site of injury, and also at remote sites. Uptake was at the cellular level, with accumulation of DDC in the cytoplasm. Cells manifesting DDC uptake were confirmed as apoptotic cells by detection of the characteristic apoptotic DNA fragmentation (positive TUNEL staining) and detection of activated caspases. The damaged region stained by DDC fluorescence correlated with the severity of neuronal deficits. Our study confirms the role of apoptosis in TBI, and proposes DDC as a useful tool for its selective targeting and detection in vivo. Such imaging of apoptosis, following future radiolabeling of DDC, may advance care for patients with head injury, by allowing real-time evaluation of the extent of tissue damage, assessment of novel therapeutic strategies, and optimization of treatment for the individual patient.